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Mycobacterium tuberculosis survival and biofilm formation studies: effect of d-amino acids, d-cycloserine and its components

The Journal of Antibiotics volume 75, pages 472–479 (2022)Cite this article

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Abstract

d-amino acids play an important role in cell wall peptidoglycan biosynthesis. Mycobacterium tuberculosis d-amino acid oxidase deletion led to reduced biofilm-forming ability. Other recent studies also suggest that the accumulation of d-amino acids blocks biofilm formation and could also disperse pre-formed biofilm. Biofilms are communities of bacterial cells protected by extracellular matrix and harbor drug-tolerant as well as persistent bacteria. In Mycobacterium tuberculosis, biofilm formation or its inhibition by d-amino acids is yet to be tested. In the present study, we used selected d-amino acids to study their role in the prevention of biofilm formation and also if d-cycloserine’s activity was due to presence of d-Serine as a metabolite. It was observed that d-serine limits biofilm formation in Mycobacterium tuberculosis H37Ra (Mtb-Ra), but it shows no effect on pre-formed biofilm. Also, d-cycloserine and its metabolic product, hydroxylamine, individually and in combination, with d-Serine, limit biofilm formation in Mtb-Ra and also disrupts existing biofilm. In summary, we demonstrated that d-alanine, d-valine, d-phenylalanine, d-serine, and d-threonine had no disruptive effect on pre-formed biofilm of Mtb-Ra, either individually or in combination, and d-cycloserine and its metabolite hydroxylamine have potent anti-biofilm activity.

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Acknowledgements

The authors would like to acknowledge funding support from SERB (Grant no. EMR/20l7/001295) and CSIR-CDRI (Grant no. MLP2033). RK is a recipient of JRF from DBT, New Delhi, India. NS, AC and MK were supported by JRF from UGC, New Delhi, India. The LC-MS/MS studies were performed at Pharmaceutics and Pharmacokinetics division of CSIR-CDRI. This manuscript is CSIR-CDRI communication no. 10404.

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Author notes

  1. These authors contributed equally: Nirbhay Singh, Anu Chauhan

Authors and Affiliations

  1. Molecular Microbiology and Immunology Division, CSIR-Central Drug Research Institute, B.S. 10/1, Sector-10, Jankipuram Extension, Sitapur Road, Lucknow, 226031, India

    Ram Kumar, Nirbhay Singh, Anu Chauhan & Sudheer Kumar Singh

  2. Jawaharlal Nehru University, New Mehrauli Road, JNU Ring Rd, New Delhi, 110067, India

    Ram Kumar, Nirbhay Singh & Mukesh Kumar

  3. Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, 201002, India

    Anu Chauhan & Sudheer Kumar Singh

  4. Pharmaceutics and Pharmacokinetics Division, CSIR-Central Drug Research Institute, B.S. 10/1, Sector-10, Jankipuram Extension, Sitapur Road, Lucknow, 226031, India

    Mukesh Kumar & Rabi Sankar Bhatta

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Contributions

SKS did the planning of experiments and analyzed the data. RK performed most of the experiments, prepared figures and did major writing work. NS and AC contributed to survival studies. MK and RSB contributed to LC-MS/MS studies. All the authors reviewed the manuscript.

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Correspondence to Sudheer Kumar Singh.

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Kumar, R., Singh, N., Chauhan, A. et al. Mycobacterium tuberculosis survival and biofilm formation studies: effect of d-amino acids, d-cycloserine and its components. J Antibiot 75, 472–479 (2022). https://doi.org/10.1038/s41429-022-00534-6

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  • DOI: https://doi.org/10.1038/s41429-022-00534-6

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